Description

• B-Mrc1-EGFP-DTR-Luciferase mice have the endogenous mouse Mrc1 promoter to determine the expression of DTR (Diphtheria toxin receptor), EGFP (enhanced green fluorescent protein) and luciferase.
• Mrc1+ macrophages cells were traced by EGFP in B-Mrc1-EGFP-DTR-Luciferase mice, and could be effectively eliminated after DT administration.

Frequency of leukocyte subpopulations in spleen

Frequency of leukocyte subpopulations in spleen by flow cytometry. Splenocytes were isolated from wild-type C57BL/6JNifdc mice (female, n=3, 6-7-week-old) and heterozygous B-Mrc1-EGFP-DTR-Luc mice (female, n=3, 6-7-week-old). Flow cytometry analysis of the splenocytes was performed to assess the frequency of leukocyte subpopulations. Percentages of T cells, B cells, NK cells, dendritic cells, monocytes, macrophages, and granulocytes in B-Mrc1-EGFP-DTR-Luc mice were similar to those in C57BL/6JNifdc mice. Values are expressed as mean ± SEM. Significance was determined by two-way ANOVA test.  *P < 0.05, **P < 0.01, ***p < 0.001.

MRC1+  cells depletion analysis in Peritoneal lavage fluid

Frequencies of MRC1+ cells in peritoneal lavage fluid by flow cytometry. Peritoneal lavage fluid was isolated from wild-type C57BL/6JNifdc mice (+/+) and heterozygous B-Mrc1-EGFP-DTR-Luciferase mice (Mut/+) injected with DT (20 ng per g body weight) for one day. Flow cytometry analysis of the peritoneal lavage fluid was performed to assess the frequencies of MRC1+ cells in CD11b-low cells, large peritoneal macrophages (LPMs) and small peritoneal macrophages (SPMs). The signal for MRC1+ cells in CD11b-low cells (A), SPMs (B) and LPMs (C) were significant decreased in heterozygous mice injected with DT.

EGFP+ depletion analysis in Peritoneal lavage fluid

Frequencies of EGFP cells in peritoneal lavage fluid by flow cytometry. Peritoneal lavage fluid was isolated from wild-type C57BL/6JNifdc mice (+/+) and heterozygous B-Mrc1-EGFP-DTR-Luciferase mice (Mut/+) injected with DT (20 ng per g body weight) for one day. Flow cytometry analysis of the peritoneal lavage fluid was performed to assess the frequencies of EGFP+ cells in CD11b-low cells, large peritoneal macrophages (LPMs) and small peritoneal macrophages (SPMs). The signal for EGFP+ cells in CD11b-low cells (A), SPMs (B) and LPMs (C) were significant decreased in heterozygous mice injected with DT.